LOCUS       sequence-447619-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0873m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in striatal cholinergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6383)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Vargas S, Lerma MN, Taskin N, Weed N, Laird WD, Bishaw YM, Bendrick 
            JL, Gore BB, Ben-Simon Y, Opitz-Araya X, Martinez RA, Way SW, 
            Thyagarajan B, Otto S, Sanchez REA, Alexander JR, Amaya A, Amster A,
            Arbuckle J, Ayala A, Baker PM, Barcelli T, Barta S, Bertagnolli D, 
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chong P, Clark M, Colbert K, 
            Daniel S, Dawe T, Departee M, DiValentin P, Donadio NP, Dotson NI, 
            Dwivedi D, Egdorf T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, 
            Gudsnuk K, Han W, Haradon Z, Hastings S, Helback O, Ho WV, Huang C, 
            Johnson T, Jones DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, 
            Loeffler H, Lusk NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor
            J, Mich JK, Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K,
            Oster AL, Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, 
            Potekhina L, Ransford S, Ray PL, Reding M, Rette DF, Reynoldson C, 
            Rimorin C, Sigler AR, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, 
            Sevigny JP, Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats 
            B, Taormina MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford 
            A, Dee N, Mollenkopf T, Ng L, Esposito L, Kalmbach BE, Yao S, Ariza 
            J, Collman F, Mufti S, Smith K, Waters J, Ersing I, Patrick M, Zeng 
            H, Lein ES, Kojima Y, Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic 
            B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   Neuron. 2025 May 21;113(10):1507-1524.e17. doi: 
            10.1016/j.neuron.2025.04.035.
  PUBMED    40403704
REFERENCE   2  (bases 1 to 6383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1507-1524.e17"
COMMENT     Sequence Label: AiP15820 -
            pAAV-AiE0873m_3xC2-minBG-RiboL1-jGC...
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1132..1149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1431..1452
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1558..1579)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1819..1838)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3210..3230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3417..3528
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3646..3786
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3646..3775
                     /label=AAV2 ITR
     rep_origin      3861..4316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3948..3967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4158..4179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4333..4352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4452..4474
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4512..4530)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4598..4702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4703..4771,4772..5563)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4921..4940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5734..6322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6223..6242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcatgga tttctccaaa gctttgtagg gcagaactct
      181 gaatgtttat ttctctgatg aatccttaat gaaagtggcc agccatcctg gggagaacaa
      241 gccttagttc ccactgagaa ggatagcaag cagcataaga gtagctgatg tcaaaggcta
      301 ttaatgcaaa tgaaaattat aattcctcat caagatctgc tcccagacag gctaatcatt
      361 gtgccacctg tttacagaaa cacactgcta atcagtcagt aaaatctttt aagtttctgt
      421 gcttctctgt gtacgatttt tagcctccat ggatttctcc aaagctttgt agggcagaac
      481 tctgaatgtt tatttctctg atgaatcctt aatgaaagtg gccagccatc ctggggagaa
      541 caagccttag ttcccactga gaaggatagc aagcagcata agagtagctg atgtcaaagg
      601 ctattaatgc aaatgaaaat tataattcct catcaagatc tgctcccaga caggctaatc
      661 attgtgccac ctgtttacag aaacacactg ctaatcagtc agtaaaatct tttaagtttc
      721 tgtgcttctc tgtgtacgat ttttagcctc catggatttc tccaaagctt tgtagggcag
      781 aactctgaat gtttatttct ctgatgaatc cttaatgaaa gtggccagcc atcctgggga
      841 gaacaagcct tagttcccac tgagaaggat agcaagcagc ataagagtag ctgatgtcaa
      901 aggctattaa tgcaaatgaa aattataatt cctcatcaag atctgctccc agacaggcta
      961 atcattgtgc cacctgttta cagaaacaca ctgctaatca gtcagtaaaa tcttttaagt
     1021 ttctgtgctt ctctgtgtac gatttttagc ctcgagctcg ggctgggcat aaaagtcagg
     1081 gcagagccat ctattgctta catttgcttc tgggatccac ccgccaccat gcatcatcac
     1141 catcatcaca cgcgtcgcaa gaagaccttc aaggaggtgg ccaccgccgt gaagatcatc
     1201 gccatgctga tgggcctgaa gatcaacgtc tatatcaagg ccgacaagca gaagaacggc
     1261 atcaaggcga acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac
     1321 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1381 ctgagcgtgg agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     1441 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1501 accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1561 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc
     1621 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1681 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1741 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag
     1801 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1861 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1921 atcctggggc acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca
     1981 gaatacaaag aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag
     2041 gagctgggga cggtgatgcg gtctctcgga cacaacccca cagaagcaga gctgcaggac
     2101 atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca
     2161 atgatggcaa gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt
     2221 gtgtttgata aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca
     2281 aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc
     2341 gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaaggggggc
     2401 agcggtgggt ctggagggac tgggggtagc ggtggtagcg gtgggacagg gggaagcggc
     2461 ggtagtggtg gaacaagcag caaagtctca cgcgacaccc tgtacgaggc ggtgcgggaa
     2521 gtcctgcacg ggaaccagcg caagcgccgc aagtttctgg agacggtgga gctgcagatc
     2581 agcctgaaga actacgaccc tcagaaggac aaacgtttct cgggcaccgt caggctcaag
     2641 tccaccccac gccccaagtt ctcggtgtgc gttctggggg accagcagca ctgtgatgaa
     2701 gccaaggccg tggatatccc ccacatggac atcgaggcgc tcaagaaact taacaaaaac
     2761 aagaagttgg tcaagaagct ggctaagaag tacgatgcct ttttggcctc tgagtctctg
     2821 attaagcaga tcccacgtat cctgggccca ggcctaaaca aggctggcaa gttcccctcc
     2881 ctgctgacac acaatgaaaa catggtggcc aaagtggatg aggtgaaatc gacaatcaag
     2941 ttccagatga agaaggtgct gtgtttggcc gtcgctgttg gccacgtgaa gatgaccgat
     3001 gatgagctag tctacaacat tcatctggct gtcaatttct tggtgtcctt gcttaagaaa
     3061 aactggcaaa acgtgcgggc tctgtacatc aagagcacca tgggcaagcc ccagcgtctg
     3121 tattagggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga ttacaaaatt
     3181 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3241 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3301 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3361 tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat cttcgactgt
     3421 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3481 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3541 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3601 agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3661 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3721 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3781 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3841 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3901 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3961 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4021 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4081 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4141 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4201 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4261 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4321 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4381 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4441 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4501 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4561 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4921 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5221 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5341 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5401 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5461 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5521 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5581 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5641 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5701 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5761 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5821 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5881 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5941 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6001 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6061 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6121 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6181 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6241 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6301 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6361 ttgctggcct tttgctcaca tgt
//
